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Science 16 June 1989:
Vol. 244. no. 4910, pp. 1307 - 1312
DOI: 10.1126/science.2660262
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Articles

Science, Vol 244, Issue 4910, 1307-1312
Copyright © 1989 by American Association for the Advancement of Science

articles

Construction of large DNA segments in Escherichia coli

M O'Connor, M Peifer, and W Bender

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.

Recombinant DNA clones containing large pieces of DNA are useful in the study of large genetic units, but these are difficult to make in most bacterial cloning vectors. A strategy is described that uses general and site-specific recombination to construct large pieces of eukaryotic DNA from smaller cloned segments. The large clones are propagated on F factor-based plasmids in Escherichia coli. They can be easily modified to introduce mutations or rearrangements. These techniques were applied to the construction of large DNA segments from the bithorax complex of Drosophila.


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